Gas Chromatography-Mass Spectrometry Detection of Thymoquinone in Oil and Serum for Clinical Pharmacokinetic Studies.

Thymoquinone (TQ) is the primary component of Nigella sativa L. (NS) oil, which is renowned for its potent hepatoprotective effects attributed to its antioxidant, anti-fibrotic, anti-inflammatory, anti-carcinogenic, and both anti- and pro-apoptotic properties. The aim of this work was to establish a method of measuring TQ in serum in order to investigate the pharmacokinetics of TQ prior to a targeted therapeutic application. In the first step, a gas chromatography-mass spectrometry method for the detection and quantification of TQ in an oily matrix was established and validated according to European Medicines Agency (EMA) criteria. For the assessment of the clinical application, TQ concentrations in 19 oil preparations were determined. Second, two serum samples were spiked with TQ to determine the TQ concentration after deproteinization using toluene. Third, one healthy volunteer ingested 1 g and another one 3 g of a highly concentrated NS oil 30 and 60 min prior to blood sampling for the determination of serum TQ level. After the successful establishment and validation of the measurement method, the highest concentration of TQ (36.56 g/L) was found for a bottled NS oil product (No. 1). Since a capsule is more suitable for oral administration, the product with the third highest TQ concentration (No. 3: 24.39 g/L) was used for all further tests. In the serum samples spiked with TQ, the TQ concentration was reliably detectable in a range between 5 and 10 µg/mL. After oral intake of NS oil (No. 3), however, TQ and/or its derivatives were not detectable in human serum. This discrepancy in detecting TQ after spiking serum or following oral ingestion may be attributed to the instability of TQ in biomatrices as well as its strong protein binding properties. A pharmacokinetics study was therefore not viable. Studies on isotopically labeled TQ in an animal model are necessary to study the pharmacokinetics of TQ using alternative modalities.

Endoscopic Resection Without Subsequent Ablation Therapy for Early Barrett's Neoplasia: Endoscopic Findings and Long-Term Mortality.

INTRODUCTION: After endoscopic resection (ER) of neoplasia in Barrett's esophagus (BE), it is recommended to ablate the remaining BE to minimize the risk for metachronous disease. However, we report long-term outcomes for a nationwide cohort of all patients who did not undergo ablation of the remaining BE after ER for early BE neoplasia, due to clinical reasons or performance status. METHODS: Endoscopic therapy for BE neoplasia in the Netherlands is centralized in 8 expert centers with specifically trained endoscopists and pathologists. Uniformity is ensured by a joint protocol and regular group meetings. We report all patients who underwent ER for a neoplastic lesion between 2008 and 2018, without further ablation therapy. Outcomes include progression during endoscopic FU and all-cause mortality. RESULTS: Ninety-four patients were included with mean age 74 (± 10) years. ER was performed for low-grade dysplasia (LGD) (10%), high-grade dysplasia (HGD) (25%), or low-risk esophageal adenocarcinoma (EAC) (65%). No additional ablation was performed for several reasons; in 73 patients (78%), the main argument was expected limited life expectancy. Median C2M5 BE persisted after ER, and during median 21 months (IQR 11-51) with 4 endoscopies per patient, no patient progressed to advanced cancer. Seventeen patients (18%) developed HGD/EAC: all were curatively treated endoscopically. In total, 29/73 patients (40%) with expected limited life expectancy died due to unrelated causes during FU, none of EAC. CONCLUSION: In selected patients, ER monotherapy with endoscopic surveillance of the residual BE is a valid alternative to eradication therapy with ablation.

Embryological signaling pathways in Barrett's metaplasia development and malignant transformation; mechanisms and therapeutic opportunities.

Barrett's metaplasia of the esophagus (BE) is the precursor lesion of esophageal adenocarcinoma (EAC), a deadly disease with a 5-year overall survival of less than 20%. The molecular mechanisms of BE development and its transformation to EAC are poorly understood and current surveillance and treatment strategies are of limited efficacy. Increasing evidence suggests that aberrant signaling through pathways active in the embryological development of the esophagus contributes to BE development and progression to EAC. We discuss the role that the Bone morphogenetic protein, Hedgehog, Wingless-Type MMTV Integration Site Family (WNT) and Retinoic acid signaling pathways play during embryological development of the esophagus and their contribution to BE development and malignant transformation. Modulation of these pathways provides new therapeutic opportunities. By integrating findings in developmental biology with those from translational research and clinical trials, this review provides a platform for future studies aimed at improving current management of BE and EAC.

An upper gastrointestinal ulcer still bleeding after endoscopy: what comes next?

INTRODUCTION: Recurrent bleeding from an upper gastrointestinal ulcer when endoscopy fails is a reason for radiological or surgical treatment, both of which have their advantages and disadvantages. CASE: Based on a patient with recurrent gastrointestinal bleeding, we reviewed the available evidence regarding the efficacy and safety of surgical treatment and embolisation, respectively. DISCUSSION: Transarterial embolisation (TAE) and surgical treatment are both options for recurrent gastrointestinal bleeding when endoscopy fails. Both therapies have serious complications and a risk of rebleeding. Choosing the therapy depends on the capability of the patient to tolerate haemodynamic instability, resuscitation and hypotension. CONCLUSION: Choosing between TAE and surgery depends a great deal on the case presented, haemodynamic stability and the skills and tools available at that moment.

Learning to perform endoscopic resection of esophageal neoplasia is associated with significant complications even within a structured training program.

BACKGROUND AND STUDY AIMS: Endoscopic resection is the cornerstone of endoscopic treatment of esophageal high grade dysplasia or early cancer. Endoscopic resection is, however, a technically demanding procedure, which requires training and expertise. The aim of the current study was to prospectively evaluate efficacy and safety of the first 120 endoscopic resection procedures of early esophageal neoplasia performed by six endoscopists (20 endoscopic resections each) who were participating in an endoscopic resection training program. PATIENTS AND METHODS: The program consisted of four tri-monthly 1-day courses with lectures, live-demonstrations, hands-on training on anesthetized pigs, and one-on-one hands-on training days. Gastroenterologists from centers with multidisciplinary expertise in upper gastrointestinal oncology participated, together with an endoscopy nurse and a pathologist. Outcome measures were complete endoscopic removal of the target area and acute complications. RESULTS: A total of 120 consecutive esophageal endoscopic resection procedures (85 ER-cap, 35 multiband mucosectomy [MBM]) were performed by six endoscopists: 109 in Barrett's esophagus, 11 for squamous neoplasia; 85 piecemeal endoscopic resections (median 3 specimens, interquartile range 2 - 4 specimens). Complete endoscopic removal was achieved in 111 /120 cases (92.5 %). Six perforations occurred (5.0 %): five were effectively treated endoscopically (clips, covered stent), and one patient underwent esophagectomy. There were 11 acute mild bleedings (9.2 %), which were managed endoscopically. Perforations occurred in ER-cap procedures performed by four participants (7.1 % ER-cap vs. 0 % MBM; P = 0.18), and in 1.7 % of the first 10 endoscopic resections and 8.3 % of the second 10 endoscopic resections per endoscopist (P = 0.26). CONCLUSION: In this intense, structured training program, the first 120 esophageal endoscopic resections performed by six participants were associated with a 5.0 % perforation rate. Although perforations were adequately managed, performing 20 endoscopic resections may not be sufficient to reach the peak of the learning curve in endoscopic resection.

Observer agreement in the assessment of narrowband imaging system surface patterns in Barrett's esophagus: a multicenter study.

BACKGROUND AND STUDY AIMS: The clinical utility of narrow-band imaging (NBI) for Barrett's esophagus is limited by the multiplicity of classification schemes. We evaluated the interobserver agreement and accuracy of a new consensus-driven simplified binary classification of NBI surface patterns.

Evaluation of management of desmoid tumours associated with familial adenomatous polyposis in Dutch patients.

BACKGROUND: The optimal treatment of desmoid tumours is controversial. We evaluated desmoid management in Dutch familial adenomatous polyposis (FAP) patients. METHODS: Seventy-eight FAP patients with desmoids were identified from the Dutch Polyposis Registry. Data on desmoid morphology, management, and outcome were analysed retrospectively. Progression-free survival (PFS) rates and final outcome were compared for surgical vs non-surgical treatment, for intra-abdominal and extra-abdominal desmoids separately. Also, pharmacological treatment was evaluated for all desmoids. RESULTS: Median follow-up was 8 years. For intra-abdominal desmoids (n=62), PFS rates at 10 years of follow-up were comparable after surgical and non-surgical treatment (33% and 49%, respectively, P=0.163). None of these desmoids could be removed entirely. Eventually, one fifth died from desmoid disease. Most extra-abdominal and abdominal wall desmoids were treated surgically with a PFS rate of 63% and no deaths from desmoid disease. Comparison between NSAID and anti-estrogen treatment showed comparable outcomes. Four of the 10 patients who received chemotherapy had stabilisation of tumour growth, all after doxorubicin combination therapy. CONCLUSION: For intra-abdominal desmoids, a conservative approach and surgery showed comparable outcomes. For extra-abdominal and abdominal wall desmoids, surgery seemed appropriate. Different pharmacological therapies showed comparable outcomes. If chemotherapy was given for progressively growing intra-abdominal desmoids, most favourable outcomes occurred after combinations including doxorubicin.

Is bacteriologic surveillance in endoscope reprocessing stringent enough?

Endoscopes, including duodenoscopes, are medical devices that are frequently associated with outbreaks of nosocomial infections. We investigated an outbreak of multidrug-resistant PSEUDOMONAS AERUGINOSA sepsis affecting three patients after endoscopic retrograde cholangiopancreaticography (ERCP). Epidemiologic investigation supplemented by molecular typing revealed that one ERCP scope was the source of infection with P. AERUGINOSA. No contamination with this microorganism was found after screening of washer-disinfectors, connecting tubes, and environmental surfaces in the endoscopy center. PSEUDOMONAS isolates from blood and endoscope channels before gas sterilization with ethylene oxide (ETO) were characterized by molecular typing as "linked isolates". Though the current surveillance system did not prevent the infections in three patients, our microbiological surveillance protocol with routine culturing of endoscopes was helpful in detecting the source of contamination and probably avoided numerous cross-contaminations in other patients who underwent ERCP procedures with endoscopes.

A systematic comparison of four different workup procedures for systematic toxicological analysis of urine samples using gas chromatography-mass spectrometry.

Sample preparation for systematic toxicological screening analysis (STA) in urine by gas chromatography-mass spectrometry (GC-MS) generally involves cleavage of conjugates by acid hydrolysis (Hy) or enzymatic hydrolysis (Gluc) followed by liquid-liquid extraction (LLE) or solid-phase extraction (SPE), and derivatization, e.g., acetylation (Ac). LLE and derivatization can be performed simultaneously, e.g., in extractive methylation (ExMe). The work presented consisted of two separate studies. In study I, 350 urine samples from 168 inpatients from an internal medicine ward were worked up by Hy-LLE-Ac, the standard workup in the authors' laboratory, Gluc-SPE-Ac, and Gluc-ExMe. In study II, 100 urine samples from psychiatric inpatients were worked up by Hy-LLE-Ac and Hy-SPE-Ac. The samples prepared were analyzed by full-scan GC-MS, and the drugs and/or their metabolites/artifacts detected after the different workup procedures were compared. The results obtained after Hy-LLE-Ac and Gluc-SPE-Ac showed only little differences, e.g., salicylic acid not being detectable with the latter. Hy-SPE-Ac covered a similar range of analytes as Hy-LLE-Ac but was much more time-consuming. Comparison of Hy-LLE-Ac and Gluc-ExMe showed that the former was better suited for basic drugs and the latter for acidic drugs, but the overlap was considerable. In conclusion, Hy-LLE-Ac remains the method of choice for STA in clinical toxicology owing to its wide analyte spectrum and short workup time. Gluc-ExMe is an ideal complementary method when acidics need to be covered. Gluc-SPE-Ac can be used as an alternative to Hy-LLE-Ac when turnaround is not critical or when automated analysis is required.

Endoscope disinfection and its pitfalls--requirement for retrograde surveillance cultures.

BACKGROUND AND STUDY AIMS: Several endoscopy-related outbreaks of infection have been reported in recent years. For early recognition of inadequate disinfection of endoscopes we designed a microbiological surveillance system to evaluate the efficacy of the cleaning and disinfection procedure, and to trace disinfection problems to individual endoscopes or washer-disinfectors. METHODS: Our surveillance protocol included anterograde and retrograde sampling, a decision algorithm, genetic fingerprinting, and scanning electron microscopy. RESULTS: Over a period of 29 months we found an increasing number of patient-ready endoscopes testing positive for Candida species other than albicans, especially C. parapsilosis. These yeasts were also isolated from the washer-disinfectors. The number of positive tests for Candida species varied from 1 out of 21 to 14 out of 27 samples from nine frequently used endoscopes. The number of colony-forming units per milliliter ranged from 1 - 10 to 3000 for endoscopes and 0.002 to 0.06 for the washer disinfectors. DNA fingerprinting was not able to discriminate different strains within C. parapsilosis. CONCLUSIONS: Our protocol was able to detect a structural problem in the endoscope disinfection process. Retrograde sampling was crucial for this purpose, because it has much higher sensitivity than anterograde sampling. Endoscopes with damaged working channels are probably the source of the contamination problem with Candida species.

[Current surveillance policy for Barrett's oesophagus in the Netherlands]. / Huidig beleid bij de surveillance van de barrettslokdarm in Nederland.

OBJECTIVE: To gain more insight into current surveillance and treatment of patients with Barrett's oesophagus with the aim of developing new guidelines. DESIGN: Questionnaire. METHOD: In 2004, a questionnaire was sent to 337 physician-endoscopists who were all registered with the Netherlands Society of Gastroenterology. The questionnaire inventoried various aspects of surveillance and treatment of patients with Barrett's oesophagus. RESULTS: Of the 289 respondents (86%), 96% carried out surveillance or had it carried out, on at least a proportion of their patients with Barrett's oesophagus. A total of 258 respondents (89%) carried out the surveillance themselves. An endoscopic indication of the presence of Barrett's oesophagus was, for 31% of the respondents, enough reason to carry out surveillance of this condition irrespective of the results of pathological investigations. 75% applied an age limit for surveillance for Barrett's. The median age limit is 75 years (interquartile distance: 70-75) and 46% of the treating professionals limited themselves to patients who, on the basis of age and co-morbidity, may undergo oesophageal resection. The choice of treatment in early neoplasia, surgical or endoscopic, depends not only on the histological diagnosis, but also on the age and the co-morbidities of the patient. CONCLUSION: Surveillance of Barrett's oesophagus is widespread in the Netherlands, and in general is carried out in accordance with international guidelines. The possibilities of treating patients with high-grade dysplasia or intramucosal carcinoma of the oesophagus endoscopically, and of consulting external advisory bodies are still insufficiently utilized.

Indications for percutaneous endoscopic gastrostomy in complex tuberculosis patients.

OBJECTIVES: Percutaneous endoscopic gastrostomy (PEG) is a widely used method for providing long-term administration of tube feeding. Different indications in relation to diseases, impairments and nutritional factors for PEG placement have been mentioned in guidelines. Treatment with PEG has not been described previously for tuberculosis (TB). Our aim was to identify and describe indications and contributing factors for PEG placement in TB patients. METHODS: A retrospective medical record review was conducted of 32 TB patients who required PEG from March 1996 to April 2004. Indications and contributing factors for PEG placement were analysed. RESULTS: PEG placement was based on three different indications. In 18 patients, PEG was used to administer tube feeding, in 4 patients anti-tuberculosis drugs were administered and in 10 patients both tube feeding and antituberculosis drugs were administered. Contributing factors for PEG placement were swallowing disabilities, weakness, anti-tuberculosis drugs and their side effects, pain of neuralgic origin, hiccups and refusal of food and drugs. CONCLUSIONS: In TB, imminent and overt malnutrition, as well as the administration of drugs with a curative aim, are new indications for PEG placement. The use of PEG can overcome various problems in TB treatment and prevent treatment default.

[Endoluminal treatment of gastro-oesophageal reflux disease: too early to determine a clear role]. / Endoscopische behandeling van gastro-oesofageale refluxziekte: nog te vroeg voor een duidelijke plaatsbepaling.

Gastro-oesophageal reflux disease is a chronic disorder. The most important therapeutic measure is long-term pharmacological acid inhibition. Anti-reflux surgery plays a role in cases where pharmacological treatment fails. A new development is endoluminal treatment for reflux disorders. The various methods used are based on one of the following principles: radiofrequency ablation, gastroplication and implantation of a bulking agent. Short-term results with endoluminal treatment are not very positive and longer-term results have been disappointing so far. Results from randomised studies are still largely unavailable. Longer follow-up and controlled randomised studies are needed before a well-founded judgment can be made regarding the place of endoluminal treatment in the therapeutic armamentarium.

Severe valproic acid intoxication is associated with atrial tachycardia: secondary detoxication by hemoperfusion.

Valproic acid is an anticonvulsant drug which is associated with serious toxicity including fatal outcome in case of severe intoxication. Secondary detoxication by hemodialysis or hemoperfusion has been employed successfully in valproic acid intoxication. Cardiac arrhythmias have only been described rarely in valproic acid intoxication in humans. We report on a 15 year-old boy with severe valproic acid intoxication (valproic acid plasma level on admission: 1 150 mg/l) who presented with coma, hypernatremia and atrial tachycardia. The patient was successfully treated with hemoperfusion and intensive supportive care without implementation of a specific antiarrhythmic therapy. We conclude that patients with severe valproic acid intoxication may benefit from secondary detoxication. In addition to generally known symptoms valproic acid intoxication may also be associated with cardiac arrhythmias.

Increased incidence of azathioprine-induced pancreatitis in Crohn's disease compared with other diseases.

BACKGROUND: Azathioprine is widely used in Crohn's disease. A major drawback is the occurrence of side-effects, especially acute pancreatitis. Acute pancreatitis is rarely seen when azathioprine is used for other diseases than Crohn's disease. AIM: To survey side-effects of azathioprine after liver or renal transplantation, for systemic lupus erythematosis, Wegener's granulomatosis, autoimmune hepatitis, rheumatoid arthritis, ulcerative colitis or Crohn's disease. METHODS: A computerized search using the term 'azathioprine' or 'imuran' was performed on the Hospital Information System of the university hospital Groningen, resulting in 1564 patients matching our criteria. RESULTS: Eleven of 224 patients with Crohn's disease experienced acute pancreatitis (4.9%) compared with two of 129 (1.5%) with autoimmune hepatitis, two of 388 (0.5%) after renal transplantation, one of 254 (0.4%) after liver transplantation. Acute pancreatitis was more prevalent in Crohn's disease compared with any other disease. Azathioprine-toxicity necessitating withdrawal occurred significantly (P < 0,05) more in rheumatoid arthritis (78 of 317), ulcerative colitis (20 of 94) and Crohn's disease (52 of 224) compared with systemic lupus erythematosis (five of 73), Wegener's granulomatosis (six of 85), autoimmune hepatitis (eight of 129), after liver transplantation (17 of 254) and after renal transplantation (22 of 388). CONCLUSIONS: Acute pancreatitis is strongly associated with Crohn's disease and rarely occurs with other underlying conditions. Overall azathioprine-induced toxicity and the necessity of withdrawal is more common in inflammatory bowel disease and rheumatoid arthritis compared with other diseases.

Concentrations and ratios of amphetamine, methamphetamine, MDA, MDMA, and MDEA enantiomers determined in plasma samples from clinical toxicology and driving under the influence of drugs cases by GC-NICI-MS.

Enantiomers of amphetamine (AM), methamphetamine (MA), 3,4-methylenedioxyamphetamine (MDA), 3,4-methylenedioxymethamphetamine (MDMA), and 3,4-methylenedioxyethylamphetamine (MDEA) exhibit different pharmacological properties. This may be important for the interpretation of analytical results. Plasma samples were analyzed using validated negative ion chemical ionization gas chromatography-mass spectrometry procedures. The results for clinical toxicology cases, divided into screening (SCR) and intoxication (ITX) cases, and those of driving under the influence of drugs (DUID) cases were compared. The concentrations of all enantiomers, except R-(-)-MDA and R-(-)- and S-(+)-MA, in the SCR samples were lower than in ITX and DUID samples. Differences between concentrations in ITX and DUID samples were only significant for both enantiomers of AM (DUID higher). These findings suggested impairment in drugged drivers. Different enantiomer ratios (R vs. S) were found for AM between DUID and SCR samples, for MDMA between ITX and SCR samples, and for MDA between DUID and ITX and DUID and SCR samples. Higher MDMA enantiomer ratios in SCR compared to ITX samples are in accordance with a previously described increase of those ratios over time, possibly allowing differentiation of recent from nonrecent ingestion. Pharmacokinetic analysis of a MDMA poisoning yielded elimination half-lives of 6.0 h for R-(-)-MDMA and 4.1 h for S-(+)-MDMA. The enantiomer ratios rose exponentially over time.

[Abdominal complaints and neurological symptoms as an early sign of lung cancer: a manifestation of the anti-Hu syndrome]. / Buikklachten en neurologische symptomen als vroege manifestatie van longkanker: een uiting van het anti-Hu-syndroom.

In two patients, women aged 73 and 46 years, gastrointestinal symptoms were initially not recognised as a paraneoplastic syndrome due to small-cell lung cancer. This led to redundant diagnostics as well as a delay in final diagnosis. The anti-Hu syndrome is characterised by the presence of anti-Hu antibodies and neurological symptoms. About a quarter of the patients with the anti-Hu syndrome will develop gastrointestinal motility disorders in the course of their illness. The primary tumour is usually a small-cell lung cancer. Whereas the presence of anti-Hu antibodies appears to be beneficial for the oncological prognosis, the neurological outcome is less favourable.

Prevalence of illicit drug use in plasmapheresis donors.

BACKGROUND AND OBJECTIVES: No data are presently available concerning the frequency of illicit drug use in plasmapheresis donors. We therefore examined source plasma units produced in the United States (US) and in Germany for evidence of illicit drug use among donors. MATERIALS AND METHODS: Seventy-five US plasma units from 10 different US states and 75 German plasma units that had been analysed principally for their protein composition were screened for drugs. Determinations were made, using automated immunoassays, of the presence of cannabis, cocaine, amphetamine, methamphetamine, methylenedioxymethamphetamine (MDMA), methylenedioxyethylamphetamine (MDE) and opiates. Positive results were confirmed by gas chromatography-mass spectrometry. RESULTS: Eleven US plasma units were found to be positive for cocaine (14.6%), whereas all German samples were cocaine negative (P = 0.0007). Fifteen US plasma units (20%) and one German unit (1.3%) were confirmed as positive for cannabis (P = 0.0003). Three out of 75 US plasma units were positive for both cannabis and cocaine. In none of the 150 samples were amphetamine, methamphetamine, MDMA, MDE or opiates detected. CONCLUSIONS: Our results strongly suggest differences in cocaine and cannabis consumption between US and German plasmapheresis donors. If these results are confirmed by larger-scale studies, random drug screening (including cocaine) of donors should be implemented in order to reduce the number of drug-containing plasma units, especially in the USA.

Recurrent severe bleeding from gastrointestinal angiodysplasia in a patient with von Willebrand's disease, controlled with recombinant factor VIIa.

A patient with von Willebrand's disease had recurrent gastrointestinal bleeding from angiodysplasia, with inadequate response to von Willebrand factor substitution, medical and endoscopic treatment, and resection of affected bowel. Frequent blood transfusions were required. She started home treatment with recombinant activated factor VII (rFVIIa) at the onset of bleeding, in addition to her standard therapy. From then on, bleeds could be controlled rapidly and no more blood transfusions were needed. We conclude that rFVIIa is effective in this case of angiodysplasia and might be a therapeutic option in similar patients.